Effects of Sevoflurane on Apoptosis of Myocardial Cells in IRI Rats

Background. Cardiomyocyte apoptosis functions essentially in ischemia/reperfusion- (I/R-) induced myocardial injury. It is suggested that autophagy is widely implicated in the regulation of cell survival and death. Sevoflurane, as a largely used inhalational general anesthetic, has been shown to have a protective effect on cardiomyocytes. However, it was yet elusive on the underlying mechanisms. Aim. The objective of this study is to investigate the association of sevoflurane-mediated cardioprotective effects with autophagy regulation. Methods. An in vitro hypoxia model was established in primary cardiomyocytes from fresh myocardial tissue of the rats. The apoptosis rate of myocardial cells treated with hypoxia and treated with sevoflurane was measured. Western blot and immunocytochemical assay were used to measure the protein expression. The cell proliferation rate and cell apoptosis were measured using the MTT assay and flow cytometry, respectively. Results. The expression of apoptotic proteins including B cell lymphoma-2 (Bcl-2), CCAAT/enhancer-binding protein homologous protein (CHOP), glucose-regulated protein 78 (GRP78), and Bcl-2-associated X protein (BAX) in myocardium treated with sevoflurane was significantly lower than that in myocardium treated with hypoxia. The expression of adhesion proteins such as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin in myocardium treated with sevoflurane was higher than that in myocardium treated with hypoxia, suggesting better connectivity of the myocardium. Conclusion. Sevoflurane treatment reduced the apoptosis of myocardial cells after hypoxia treatment.

Frequency of transovarial transmission of dengue virus in Aedes aegypti mosquitoes in endemic areas of dengue hemorrhagic fever in Ceper District, Klaten Regency, Central Java Province, Indonesia

Klaten Regency is an area in Central Java Province with a high rate of endemic dengue hemorrhagic fever. This study aims to demonstrate a correlation between transovarial transmission and the incidence of dengue haemorrhagic fever in Ceper District, Klaten Regency. Aedes larvae and pupae were collected from January 2019 to March 2019 from various containers, using gamadotic and dipper tools, in Ngawonggo, Kajen, and Meger villages, which have high, medium, and low numbers of dengue cases, respectively. The larvae and pupae obtained were reared into adults. The presence of dengue virus in head squashes of 1-week-old female Ae. aegypti was identified based on an immunocytochemical assay using DSSE10 monoclonal antibody, made at Universitas Gadjah Mada as a primary antibody. The frequency of transovarial transmission of dengue virus (TIR) in Ae. aegypti in the three villages was 35.7%. The highest frequency occurred in Ngawonggo Village, at 40 %, and the lowest in Meger Village, at 28%, while the frequency in Kajen Village was 36.9 %. This study proves that there is a strong correlation (r = 0.99664) between transovarial transmission and the incidence of dengue hemorrhagic fever in Ceper District, Klaten Regency, Central Java.

Complex cytological and immunocytochemical study of cerebrospinal fluid in the intracranial breast carcinoma metastases diagnosis

Introduction. Metastases of breast cancer are detected in 5 to 48% cases, depending on the biological tumor subtype, which is the second most common after lung cancer in men. Cytological liquor analysis is one of the “gold standards” for the metastatic brain lesion identification, and, among other modern approaches, it can increase the CNS metastasis detection. Breast cancer cell identification in cerebrospinal fluid is a poor prognostic factor. A cytological examination has a predictive significance for these patients: the absence of tumor cells in the cerebrospinal fluid can be considered the sign of the intrathecal therapy effectiveness. The accuracy of the cytological diagnosis directly depends on the detailed patient’s history. Immunocytochemical assays increase the sensitivity of the cytological diagnostics. The aim of the study was to evaluate the ca-pabilities of comprehensive cytological and immunocytochemical studies in the secondary cerebral lesions diagnosis in breast cancer patients. Materials and methods. We conducted 169 cytological studies in 55 breast cancer patients with brain metas-tases. Liquor samples were obtained using lumbar punctures. 36 patients underwent 44 immunocytochemical studies and 117 immunocytochemical reactions to clarify the cytological diagnosis. Results. The sensitivity of the cytological method was 69.8%. Immunocytochemistry increased the sensitiv-ity to 72.3%. In 29 patients, the tumor histogenesis was determined using an immunocytochemical assay. Conclusion. The liquor cytological analysis is frequently the only primary morphologic diagnostic method to detect breast cancer metastasis within the central nervous system. Immunocytochemical assay significantly expands the possibilities of cytological study, allowing health care specialists to detect single tumor cells in samples and determine the metastatic neoplasm histogenesis in most cases, which also increases sensitivity. Keywords: breast cancer, cytological and immunocytochemical study of cerebrospinal fluid

Ultrastructural and immunocytochemical analysis of circulating hemocytes from Cherax quadricarinatus (von Martens, 1868)

Transmission electron microscopy (TEM) and immunocytochemistry were used to assay the circulating hemocytes characteristics in Cherax quadricarinatus (von Martens, 1868). Three haemocyte types could be identified in circulating hemocytes by TEM, i.e. hyaline cells (HC), semigranular cells (SGC), or granular cells (GC). The circulating hemocytes from C. quadricarinatus could be separated into layers by density gradient centrifugation. The CD34+ cells which are believed to be stem cell-like cells has been discovered in the first layer via immunocytochemical assay, which has not been reported in previous studies. These CD34+ cells may be immature cells derived from a single undifferentiated precursor cell line in specialized hematopoietic tissue (HPT).